Food DNA isolation miniprep kit: Manual
Food DNA isolation miniprep kit: Manual
Food DNA isolation miniprep kit: Manual
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5. Column washing 1<br />
Wash the column with 500µl Wash Buffer 1 and centrifuge at 5,000 x g for 1 min.<br />
Discard through.<br />
Ensure that ethanol has been added into the Wash Buffer 1 before use (refer to Reconstitution of<br />
Solutions).<br />
6. Column washing 2<br />
Wash the column with 500µl Wash Buffer 2 and centrifuge at 5,000 x g for 1 min.<br />
Discard Flow through. Wash the column again with 500µl Wash Buffer 2 and centrifuge<br />
at maximum speed for 3 min.<br />
Ensure that ethanol has been added into the Wash Buffer 2 before use (refer to Reconstitution of<br />
Solutions). Ensure centrifugation for 3 min to remove ethanol completely.<br />
7. <strong>DNA</strong> Elution<br />
Place the column into a clean microcentrifuge tube. Add 50-100µl of preheated Elution<br />
Buffer, TE buffer or sterile water directly onto column membrane and stand for 2 min.<br />
Centrifuge at 5,000 x g for 1 min to elute <strong>DNA</strong>. Store <strong>DNA</strong> at 4°C or -20°C.<br />
Ensure that the Elution Buffer is dispensed directly onto the center of the membrane for complete<br />
elution. TEBuffer can also elute <strong>DNA</strong> although EDTA may inhibit subsequent enzymatic<br />
reactions. If water is used for eluting <strong>DNA</strong>, maximum elution efficiency is achieved between<br />
pH7.0 and 8.5. Store <strong>DNA</strong> at -20°C as <strong>DNA</strong> may degrade in the absence of a buffering agent.<br />
Note: The recommended amplicon length for PCR analysis is